CN114544972A - 一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法 - Google Patents
一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法 Download PDFInfo
- Publication number
- CN114544972A CN114544972A CN202210171180.4A CN202210171180A CN114544972A CN 114544972 A CN114544972 A CN 114544972A CN 202210171180 A CN202210171180 A CN 202210171180A CN 114544972 A CN114544972 A CN 114544972A
- Authority
- CN
- China
- Prior art keywords
- yersinia
- intermedia
- yersinia enterocolitica
- enterocolitica
- protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 21
- 241000607734 Yersinia <bacteria> Species 0.000 title abstract description 26
- 208000010227 enterocolitis Diseases 0.000 title abstract description 12
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 25
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 25
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 claims abstract description 9
- 241000607447 Yersinia enterocolitica Species 0.000 claims description 33
- 229940098232 yersinia enterocolitica Drugs 0.000 claims description 33
- 241000607481 Yersinia intermedia Species 0.000 claims description 24
- 238000002156 mixing Methods 0.000 claims description 12
- 238000001819 mass spectrum Methods 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 238000004458 analytical method Methods 0.000 claims description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 235000010469 Glycine max Nutrition 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 210000002421 cell wall Anatomy 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 235000019253 formic acid Nutrition 0.000 claims description 3
- 239000011159 matrix material Substances 0.000 claims description 3
- 238000001906 matrix-assisted laser desorption--ionisation mass spectrometry Methods 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000007790 scraping Methods 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 239000012137 tryptone Substances 0.000 claims description 2
- NZNMSOFKMUBTKW-UHFFFAOYSA-N Cyclohexanecarboxylic acid Natural products OC(=O)C1CCCCC1 NZNMSOFKMUBTKW-UHFFFAOYSA-N 0.000 claims 1
- AFVLVVWMAFSXCK-VMPITWQZSA-N alpha-cyano-4-hydroxycinnamic acid Chemical compound OC(=O)C(\C#N)=C\C1=CC=C(O)C=C1 AFVLVVWMAFSXCK-VMPITWQZSA-N 0.000 claims 1
- 238000004949 mass spectrometry Methods 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 15
- 244000052616 bacterial pathogen Species 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 1
- 238000001228 spectrum Methods 0.000 abstract 1
- 235000013305 food Nutrition 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 4
- AFVLVVWMAFSXCK-UHFFFAOYSA-N α-cyano-4-hydroxycinnamic acid Chemical compound OC(=O)C(C#N)=CC1=CC=C(O)C=C1 AFVLVVWMAFSXCK-UHFFFAOYSA-N 0.000 description 3
- 206010035148 Plague Diseases 0.000 description 2
- 206010048249 Yersinia infections Diseases 0.000 description 2
- 208000025079 Yersinia infectious disease Diseases 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 241000143060 Americamysis bahia Species 0.000 description 1
- 241000272517 Anseriformes Species 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 241000237519 Bivalvia Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000272161 Charadriiformes Species 0.000 description 1
- 241000700114 Chinchillidae Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 241000772415 Neovison vison Species 0.000 description 1
- 241000237502 Ostreidae Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 235000020639 clam Nutrition 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000020636 oyster Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 210000004916 vomit Anatomy 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
- G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56916—Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Bioinformatics & Computational Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Electrochemistry (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法，利用基质辅助激光解析电离飞行时间质谱检测小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的肽指纹图谱，根据两种耶尔森氏菌特征峰的不同，鉴定其特征蛋白，区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌。本发明采用的方法比现有的传统生化实验鉴定时间缩短1‑2天，可以快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌，实现快速检测。该方法自动化程度高，检测时间短，有效降低一线检测人员接触食源致病菌的几率，减少食源致病菌对人和环境污染的风险。
Description
技术领域
本发明涉及一种利用基质辅助激光解析电离飞行时间质谱(Matrix-AssistedLaser Desorption Ionization Time-of-Flight Mass Spectrometry,MALDI-TOF MS)技术快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法。
背景技术
小肠结肠炎耶尔森氏菌(Yersinia enterocolitica)的分布很广，可存在于生的蔬菜、乳和乳制品、肉类、豆制品、沙拉、牡蛎、蛤和虾。也存在于环境中，如湖泊、河流、土壤和植被。已从家畜、狗、猫、山羊、灰鼠、水貂和灵长类动物的粪便中分离出该菌。在港湾周围，许多鸟类包括水禽和海鸥可能是带菌者。它能引起一种叫做耶尔森氏菌病(Yersiniosis)的人兽共患病，不但能够感染猪、牛、羊等各类动物，而且被感染的动物排出的粪便中也携带该菌，从而污染人类食用的食品、水源等资源，导致人类发生呕吐、腹泻等不适症状，严重者甚至能够威胁到生命。世界各地陆续报道的由本菌引起的败血症，其死亡率可达34％～50％。并且，由于该菌的生存条件极广，生长温度0℃～44℃，甚至报道在-2℃仍可生存，生长环境pH4.0～pH10.0，可以适应中强酸碱。因此，小肠结肠炎耶尔森氏菌是对人类身体健康具有极大威胁的一种细菌。受到小肠结肠炎耶尔森氏菌污染的食物与正常食物在外形与味觉上没有区别，因此准确、快速地检测食品中的小肠结肠炎耶尔森氏菌具有十分重要的意义。小肠结肠炎耶尔森氏菌的检测多采用传统的生化反应实验等，该方法所需时间长，一般情况下需要5-7天，很难满足快速检测的需要。
食品安全国家标准《GB 4789.8-2016食品微生物学检验小肠结肠炎耶尔森氏菌检验》中，小肠结肠炎耶尔森氏菌典型菌落在CIN-1上为深红色中心，周围具有无色透明圈(红色牛眼状菌落)，菌落大小为1mm-2mm，在改良Y琼脂平板上为无色透明、不黏稠的菌落。中间型耶尔森氏菌(Yersinia intermedia)在平板上形态与小肠结肠炎耶尔森氏菌相似，两者难以区分，生化鉴定鉴定所需时间长、实验步骤繁琐。
随着现代科学技术的不断发展，特别是免疫学、分子生物学的不断发展，人们已创建了不少快速的小肠结肠炎耶尔森氏菌检测方法。PCR技术，是一种快速、灵敏、特异性好的技术，但是目前该技术还是依赖于传统方法的前增菌步骤，增菌液中可能含有PCR抑制剂，从而影响PCR的扩增结果。小肠结肠炎耶尔森氏菌和中间型耶尔森氏菌都属于耶尔森氏菌属，文献报道耶尔森氏菌属抗原性复杂，其抗原特性与其它肠杆菌科的抗原成分相似，难以进行属种间特异性鉴别。
发明内容
本发明所要解决的技术问题是提供一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法，以克服现有技术的上述缺点，从而为食品安全提供科学的依据和指导作用。
本发明的主要原理为：利用基质辅助激光解析电离飞行时间质谱(Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry,MALDI-TOF MS)技术检测小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌蛋白图谱，根据两种菌产生的蛋白特征峰达到区分的目的，具体包括下列步骤：
1、小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的待测样品制备
接种环分别转接小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌到胰蛋白胨大豆琼脂平板上，26℃培养24～48h；刮取样品5-10mg，加入300μl水，混匀后再加入900μl无水乙醇，仔细混匀。高速离心2min，弃去上请。加入50μl 70％甲酸，仔细混匀，再加入50μl乙腈，仔细混匀，超声破壁10min，高速离心2min，吸出上清。先点1μl样品上清，放干后再点1μlCHCA(α-氰基-4-羟基肉桂酸)基质，晾干后上机进行检测。
2、MALDI-TOF MS/MS分析
参数设置：分析采用反射模式，氮气激光光源，激光频率:80Hz，质谱扫描范围:200-2000Da，每个样品激光轰击次数:100次。
3、蛋白图谱分析
质谱采集的数据通过MASCOT数据库匹配蛋白序列，NCBI网站搜索鉴定蛋白种类。
本发明的有益效果：
本发明的方法作为一种筛选工具，比现有的传统生化实验鉴定时间缩短1-2天，可以快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌，实现快速检测。相对于PCR方法和免疫方法，该筛选方法自动化程度更高，降低一线检测人员接触食源致病菌的几率，减少食源致病菌对人和环境污染的风险。
说明书附图
图1是小肠结肠炎耶尔森氏菌的部分蛋白一级质谱图。
图2是中间型耶尔森氏菌的部分蛋白一级质谱图。
图3是小肠结肠炎耶尔森氏菌的部分蛋白二级质谱图。
图4是中间型耶尔森氏菌的部分蛋白二级质谱图。
具体实施方式
下列实施例进一步说明本发明，但不应当作对本发明的限制。
实施例1
(1)小肠结肠炎耶尔森氏菌(NICPBP52203)与中间型耶尔森氏菌(CGMCC 1.6197)培养物的制备
接种环无菌操作分别转接小肠结肠炎耶尔森氏菌和中间型耶尔森氏菌到营养琼脂平板上，26℃培养24h。刮取样品5-10mg，加入300μl水，混匀后再加入900μl无水乙醇，仔细混匀。高速离心2min，弃去上请。加入50μl 70％甲酸，仔细混匀，再加入50μl乙腈，仔细混匀，超声破壁10min，高速离心2min，吸出上清。先点1μl上清，放干后再点1μl CHCA基质，晾干后上机进行检测。
(2)MALDI-TOF MS/MS分析
分析采用反射模式，氮气激光光源，激光频率:80Hz，质谱扫描范围:200-2000Da，每个样品激光轰击次数:100次。
(3)蛋白图谱分析
质谱采集的数据通过MASCOT数据库匹配蛋白序列，NCBI网站搜索鉴定蛋白种类。
结果见图1-4，图1-4是小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的部分蛋白质谱图谱。
方法验证
一种快速区分小肠结肠炎耶尔森氏菌和中间型耶尔森氏菌的检测方法，该方法对来源于泡菜等食品中的小肠结肠炎耶尔森氏菌和中间型耶尔森氏菌进行方法验证，通过比较国家标准GB 4789.8-2016和该方法，检测数据表明通过特征蛋白(DUF3560 domain-containing protein)的差异有效区分小肠结肠炎耶尔森氏菌和中间型耶尔森氏菌，方法的稳定性良好，可以应用于日常检测中区分小肠结肠炎耶尔森氏菌和中间型耶尔森氏菌。
当然，上述说明并非是对本发明的限制，本发明也并不限于上述举例，本技术领域的普通技术人员，在本发明的实质范围内，作出的变化、改型、添加或替换，都应属于本发明的保护范围。
Claims (3)
1.一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法，其特征在于，利用基质辅助激光解析电离飞行时间质谱技术检测小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的蛋白图谱，根据两种耶尔森氏菌特征峰的不同，鉴定其特征蛋白以区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌。
2.如权利要求1所述的检测方法，其特征在于，小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌二级质谱所得的特征蛋白不同。
3.如权利要求1或2所述的检测方法，其特征在于，所述的检测方法具体包括下列步骤：
（1）小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的待测样品制备
接种环分别转接小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌到胰蛋白胨大豆琼脂平板上，26 ℃培养24 h；刮取样品5-10 mg，加入300 μl水，混匀后再加入900 μl无水乙醇，仔细混匀；高速离心2 min，弃去上请；加入50 μl 70%甲酸，仔细混匀，再加入50 μl乙腈，仔细混匀，超声破壁10 min，高速离心2 min，吸出上清；先点1 μl上清，放干后再点1 μl CHCA基质，晾干后上机进行检测；
（2）MALDI-TOF MS/MS分析
参数设置：分析采用反射模式，氮气激光光源，激光频率: 80Hz，质谱扫描范围: 200-2000Da，每个样品激光轰击次数: 100 次；
（3）小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的特征蛋白区分
小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌产生的蛋白图谱不同，主要区别在某些特征峰不一致，小肠结肠炎耶尔森氏菌核质比主要在713、860、876，中间型耶尔森氏菌核质比主要在860、876；在核质比为713处，小肠结肠炎耶尔森氏菌所具有的特征蛋白为DUF3560domain-containing protein。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210171180.4A CN114544972A (zh) | 2022-02-23 | 2022-02-23 | 一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210171180.4A CN114544972A (zh) | 2022-02-23 | 2022-02-23 | 一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114544972A true CN114544972A (zh) | 2022-05-27 |
Family
ID=81677355
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210171180.4A Pending CN114544972A (zh) | 2022-02-23 | 2022-02-23 | 一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114544972A (zh) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109696546A (zh) * | 2019-02-25 | 2019-04-30 | 河南省商业科学研究所有限责任公司 | 一种检测小肠结肠炎耶尔森氏菌的方法 |
-
2022
- 2022-02-23 CN CN202210171180.4A patent/CN114544972A/zh active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109696546A (zh) * | 2019-02-25 | 2019-04-30 | 河南省商业科学研究所有限责任公司 | 一种检测小肠结肠炎耶尔森氏菌的方法 |
Non-Patent Citations (4)
| Title |
|---|
| PETER LASCH等: "Characterization of Yersinia using MALDI-TOF mass spectrometry and chemometrics", 《ANAL CHEM》, vol. 82, no. 20, 15 October 2010 (2010-10-15), pages 8464 - 8475 * |
| 王丽丽等: "北京市市售生禽肉及社区居民冰箱耶尔森菌污染状况及分子特征研究", 《中国人兽共患病学报》, vol. 36, no. 9, 30 September 2020 (2020-09-30), pages 752 - 757 * |
| 王伟等: "基质辅助激光解吸电离飞行时间质谱技术对阪崎克罗诺杆菌的鉴定及溯源", 《中国食品卫生杂志》, vol. 30, no. 2, 30 March 2018 (2018-03-30), pages 125 - 131 * |
| 王明贵: "《感染性疾病与抗微生物治疗》", 31 August 2020, 复旦大学出版社, pages: 20 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Brown et al. | Frequency and spatial distribution of environmental Campylobacter spp | |
| Singhal et al. | MALDI-TOF MS in clinical parasitology: applications, constraints and prospects | |
| James et al. | Comparison of cell culture, mouse inoculation, and PCR for detection of Toxoplasma gondii: effects of storage conditions on sensitivity | |
| Lewis | Diagnostic tests for chancroid | |
| Panangala et al. | Intra‐and interspecific phenotypic characteristics of fish‐pathogenic Edwardsiella ictaluri and E. tarda | |
| Pagmadulam et al. | Seroprevalence of Toxoplasma gondii and Neospora caninum infections in cattle in Mongolia | |
| CN105651679A (zh) | 一种基于高光谱成像技术对培养基上的细菌菌落进行快速分类的方法 | |
| Waap et al. | Prevalence and geographic distribution of Besnoitia besnoiti infection in cattle herds in Portugal | |
| Gharekhani et al. | Herd-level contamination of Neospora caninum, Toxoplasma gondii and Brucella in milk of Iranian dairy farms | |
| Mougin et al. | Rapid identification of Vibrio Species of the harveyi clade using MALDI-TOF MS profiling with main spectral profile database implemented with an in-house database: luvibase | |
| Šilha et al. | Modified isolation method of Arcobacter spp. from different environmental and food samples | |
| Kazazić et al. | Matrix-assisted laser desorption/ionization time of flight mass spectrometry identification of Vibrio (Listonella) anguillarum isolated from sea bass and sea bream | |
| Mallinson et al. | Improved plating media for the detection of Salmonella species with typical and atypical hydrogen sulfide production | |
| CN114544972A (zh) | 一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法 | |
| Ma et al. | Differentiation of fecal Escherichia coli from human, livestock, and poultry sources by rep-PCR DNA fingerprinting on the shellfish culture area of East China Sea | |
| Winters et al. | Rapid PCR with nested primers for direct detection ofCampylobacter jejuniin chicken washes | |
| Hochel et al. | Detection of Campylobacter species in foods by indirect competitive ELISA using hen and rabbit antibodies | |
| Thompson | Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry in veterinary medicine: Recent advances (2019–present) | |
| Bokma et al. | Non-specific, agar medium-related peaks can result in false positive Mycoplasma alkalescens and Mycoplasma arginini identification by MALDI-TOF MS | |
| Achakzai et al. | Microbiological studies on Clostridium perfringens isolated from commercial poultry of Balochistan | |
| Sprygin et al. | Biological characterization of Russian Mycoplasma gallisepticum field isolates | |
| Mata et al. | Comparison of conventional and rapid methods for Salmonella detection in Artisanal Minas Cheese | |
| Wu et al. | Extension and application of a database for the rapid identification of Vibrio using MALDI-TOF MS | |
| CN107988342B (zh) | 质谱鉴定副溶血性弧菌分型的方法及产品 | |
| Khan et al. | Cultural and immunological methods for the detection of Campylobacter jejuni: A review |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |